Metabolic Degradation and Bioactive Derivative Synthesis of Phenazine-1-Carboxylic Acid by Genetically Engineered Pseudomonas chlororaphis HT66

Phenazine-1-carboxylic acid (PCA) secreted by Pseudomonas chlororaphis has been commercialized and widely employed as an antifungal pesticide. However, it displays potential hazards to nontarget microorganisms and the environment. Although the PCA degradation characteristics have received extensive attention, the biodegradation efficiency is still insufficient to address the environmental risks. In this study, an engineered Pseudomonas capable of degrading PCA was constructed by introducing heterologous PCA 1,2-dioxygenase (PcaA1A2A3A4). By integrating the PCA degradation module in the chemical mutagenesis mutant P3, 7.94 g/L PCA can be degraded in 60 h, which exhibited the highest PCA degradation efficiency to date and was 35.4-fold higher than that of the PCA natural degraders. Additionally, PCA was converted to 1-methoxyphenazine through structure modification by introducing the functional enzymes PhzSPa and PhzMLa, which has good antifungal activity and environmental compatibility. This work demonstrates new possibilities for developing PCA-derived biopesticides and enables targeted control of the impact of PCA in diverse environments.